AIM: To explore the role of focal adhesion kinase (FAK) in the apoptosis in culture-activated rat hepatic stellate cells (HSCs) using a specific anti-FAK antibody. METHODS: Rat HSCs were prepared from Wistar rats by in situ perfusion of collagenase and pronase and single-step density Nycodenze gradient. Culture-activated HSCs were serum-starved and treated with the anti-FAK antibodies for 24, 48 or 72 h. The apoptosis of HSC was detected by DNA-fragment assay, flow cytometry and caspase-3 activity determination. The expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNA was assessed by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The experiment showed that anti-FAK antibodies induced apoptosis of culture-activated rat HSCs. This phenomenon displayed the classical features of apoptotic cell death (DNA fragmentation, cell cycle analysis) after treated with 30 mg.L(-1) FAK antibody for 72 h, and accompanied by a significant increase of caspase-3 activity(1208+/-76) vs (309+/-28) nmol.min(-1).g(-1), t=208.5, P<0.05. Meanwhile, treatment with the FAK antibody in HSCs could markedly decrease the TIMP-1 mRNA expression (0.07+/-0.01 vs 0.38+/-0.03, t=2.72, P<0.05). CONCLUSION: FAK plays an important role in the survival of HSCs and the specific anti-FAK antibody could induce the apoptosis in rat HSCs.
AIM: To explore the role of focal adhesion kinase (FAK) in the apoptosis in culture-activated rat hepatic stellate cells (HSCs) using a specific anti-FAK antibody. METHODS:Rat HSCs were prepared from Wistar rats by in situ perfusion of collagenase and pronase and single-step density Nycodenze gradient. Culture-activated HSCs were serum-starved and treated with the anti-FAK antibodies for 24, 48 or 72 h. The apoptosis of HSC was detected by DNA-fragment assay, flow cytometry and caspase-3 activity determination. The expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) mRNA was assessed by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The experiment showed that anti-FAK antibodies induced apoptosis of culture-activated rat HSCs. This phenomenon displayed the classical features of apoptotic cell death (DNA fragmentation, cell cycle analysis) after treated with 30 mg.L(-1) FAK antibody for 72 h, and accompanied by a significant increase of caspase-3 activity(1208+/-76) vs (309+/-28) nmol.min(-1).g(-1), t=208.5, P<0.05. Meanwhile, treatment with the FAK antibody in HSCs could markedly decrease the TIMP-1 mRNA expression (0.07+/-0.01 vs 0.38+/-0.03, t=2.72, P<0.05). CONCLUSION:FAK plays an important role in the survival of HSCs and the specific anti-FAK antibody could induce the apoptosis in rat HSCs.
Authors: Elena V Kurenova; Darell L Hunt; Dihua He; Andrew T Magis; David A Ostrov; William G Cance Journal: J Med Chem Date: 2009-08-13 Impact factor: 7.446
Authors: Elena Kurenova; Li-Hui Xu; Xihui Yang; Albert S Baldwin; Rolf J Craven; Steven K Hanks; Zheng-Gang Liu; William G Cance Journal: Mol Cell Biol Date: 2004-05 Impact factor: 4.272